Protocols

Specificity / Sensitivity

EGF Receptor(1F4) Mouse mAb detects endogenous levels of EGF receptors. It does not cross-react with other Erb family members.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a recombinant protein corresponding to the C-terminal sequence of human EGF receptor.

Western Blotting

Western blot analysis of extracts from the indicated cell lines. EGF Receptor(1F4) Mouse mAb specifically detects EGF receptors expressed in A431 and MDA-MD-468 cells, but does not cross-react with ErbB2, 3, and 4 proteins expressed in MDA-MD-453 and MCF-7 cells.

Background

The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for the adaptor protein c-Cbl, leading to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provide a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).